Peripherin and Neurofilament Light Protein Splice Variants in Amyotrophic Lateral Sclerosis (ALS)

Abstract

Nucleotide sequences encoding novel splice variants of peripherin and neurofilament light protein, proteins encoded by the novel splice variants and antibodies thereto are disclosed. In addition, methods are described for detecting ALS in a subject suspected of having ALS, comprising detecting the presence or absence of the novel splice variants or resulting proteins or a change in the amount of the novel splice variants or resulting proteins; wherein the presence or change in the amount of the nucleotide sequence is indicative of ALS.

1 . An isolated nucleic acid molecule comprising
 (a) a nucleic acid sequence as shown in SEQ ID NO:2, 27, 7, or 9, wherein T can also be U;   (b) a nucleic acid sequence that is complementary to a nucleic acid sequence of (a);   (c) a nucleic acid sequence that has substantial sequence homology to a nucleic acid sequence of (a) or (b);   (d) a nucleic acid sequence that is an analog of a nucleic acid sequence of (a), (b) or (c);   (e) a nucleic acid sequence that hybridizes to a nucleic acid sequence of (a), (b), (c) or (d) under stringent hybridization conditions; and   (f) a nucleic acid sequence differing from any of the nucleic acid sequences of (a) to (e) in codon sequences due to the degeneracy of the genetic code.   
     
     
         2 . An isolated nucleic acid molecule encoding an amino acid sequence as shown in SEQ ID NOs:5, 28, 7 or 9. 
     
     
         3 . A recombinant expression vector comprising the isolated nucleic acid molecule of  claim 1 . 
     
     
         4 . An siRNA molecule comprising the RNA sequence as shown in SEQ ID NOs: 19-22 or SEQ ID NOs: 23 and 24 or SEQ ID NOs:25 and 26. 
     
     
         5 . An isolated polypeptide comprising the amino acid sequence as shown in SEQ ID NOs:5, 28, 7 or 9 or a fragment thereof. 
     
     
         6 . The isolated polypeptide of  claim 5 , wherein the fragment comprises VSGPGIRGGF (SEQ ID NO:4). 
     
     
         7 . The isolated polypeptide of  claim 5 , wherein the fragment comprises VQEPGGPARRDAGVVSRVPAD (SEQ ID NO:29). 
     
     
         8 . The isolated polypeptide of  claim 5 , wherein the fragment comprises NDLKSIRDLR (SEQ ID NO:8). 
     
     
         9 . The isolated polypeptide of  claim 5 , wherein the fragment comprises ARKGAKNTDA (SEQ ID NO:10). 
     
     
         10 . A binding protein that binds to the isolated polypeptide of  claim 5 . 
     
     
         11 . The binding protein of  claim 10 , wherein the binding protein is an antibody or fragment thereof. 
     
     
         12 . The binding protein of  claim 11 , wherein the antibody is a monoclonal antibody. 
     
     
         13 . A method of detecting ALS in a subject suspected of having ALS, comprising detecting the presence or absence of a nucleotide sequence or a change in the amount of the nucleic acid sequence encoding the polypeptide having the amino acid sequence as shown in SEQ ID NO:5, 28, 7 or 9; wherein the presence or change in the amount of the nucleotide sequence is indicative of ALS. 
     
     
         14 . The method of  claim 13 , wherein detecting the presence or absence or change in the amount of the nucleotide sequence comprises contacting the sample under hybridization conditions with one or more nucleotide probes labeled with a detectable marker. 
     
     
         15 . A method of detecting, monitoring or diagnosing ALS in a subject comprising the steps of:
 (a) contacting a sample of said subject with the binding protein of  claim 10 ;   (b) measuring the amount of binding protein-protein complex in the sample; and   (c) comparing the amount of binding protein-protein complex in the sample to a control;   wherein a difference in the amount of binding protein-protein complex in the sample as compared to control is indicative of ALS or the stage of ALS.   
     
     
         16 . The method of  claim 15 , wherein the difference in the amount of binding protein-protein complex is an increase. 
     
     
         17 . The method of  claim 15 , wherein the difference in the amount of binding protein-protein complex is a decrease. 
     
     
         18 . The method of  claim 15 , wherein the sample comprises cerebrospinal fluid, plasma, blood serum, whole blood, spinal cord tissue, brain cells, motor neurons, urine or peripheral blood cells. 
     
     
         19 . The method of  claim 15 , wherein the subject is an animal. 
     
     
         20 . The method of  claim 19 , wherein the animal is human. 
     
     
         21 . The method of  claim 15 , further comprising assessing the subject using traditional techniques for ALS. 
     
     
         22 . A kit comprising the binding protein of  claim 10  and instructions for use. 
     
     
         23 . The kit of  claim 22 , wherein the binding protein is labeled.